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We studied a mesoporous silica nanosphere (MSN) material with tunable release capability for drug delivery applications. We employed luciferase chemiluminescence imaging to investigate the kinetics and mechanism of the adenosine 5-triphosphate (ATP) release with various disulfide-reducing agents as uncapping triggers. ATP molecules were encapsulated within the MSNs by immersing dry nanospheres in aqueous solutions of ATP followed by capping of the mesopores with chemically removable caps, such as cadmium sulfide (CdS) nanoparticles and poly(amido amine) dendrimers (PAMAM), via a disulfide linkage. By varying the chemical nature of the ''cap'' and ''trigger'' molecules in our MSN system, we discovered that the release profiles could indeed be regulated in a controllable fashion.

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We investigated G protein-stimulated release of ATP from human umbilical vein endothelial cells (HUVECs) using the G protein stimulant compound 48/80. Application of compound 48/80 resulted in dose-dependent ATP evolution from cultured HUVECs. This release was not cytotoxic as demonstrated by a lactate dehydrogenase assay and the ability of the cells to load and retain the viability dye calcein following stimulation. Mastoparan also stimulated release of ATP, further suggesting the process was G-protein initiated. This G protein was insensitive to pertussis toxin and appeared to be of the Gq-subtype. The ATP efflux was completely abolished in the presence of EGTA and thapsigargin signifying a strict Ca2+ dependence. Furthermore, compound 48/80-induced release was significantly decreased in cells pretreated with the phospholipase C inhibitor U73122. Thus, the release pathway appears to proceed through an increase in intracellular Ca2+ via PLC activation. Additionally, the G protein-initiated release was attenuated by pretreatment of the cells with either phorbol ester or indolactam V, both activators of protein kinase C. Finally, ATP release was not affected by treating HUVECs with nitric oxide synthase (NOS) inhibitors or glybenclamide.